International Refereed Journal of Engineering and Science (IRJES)

International Refereed Journal of Engineering and Science (IRJES)
ISSN (Online) 2319-183X, (Print) 2319-1821
Volume 2, Issue 5(May 2013), PP.46-53
www.irjes.com
www.irjes.com 46 | Page
Start-Up Of An Upflow Anaerobic Sludge Blanket Reactor
Treating Low-Strength Wastewater Inoculated With
Non-Granular Sludge
K.R.Venkatesh, M.Rajendran & A.Murugappan
Department of Civil Engineering, Faculty of Engineering and Technology, Annamalai University,
Annamalai Nagar- 608 002, Tamilnadu, India.
Abstract :The upflow anaerobic sludge blanket (UASB) process is known to be energy conservative
biotechnology. Its low cost and low skill requirement render it to be a viable technology for reducing organic
pollution loads. However, this system is facing a challenge in the treatment of low-strength wastewater
especially inoculated with non-granular sludge. This work was performed to emphasis an in-depth
understanding of an UASB reactor start-up process inoculated with digested slurry for treating synthetic
wastewater of chemical oxygen demand (COD) around 700-1000 mg/L. The lab scale reactor with an effective
volume of 9.97 L was operated under ambient temperature (24-350
C). At the end of start-up period that lasted
for 84 days, this system achieved 90.8% COD removal and a biogas evolution of 4.72 L/d (457 L / kg COD
removal) at an organic loading rate (OLR) of 1.293 kg COD/ m3
d. Concurrently, the volatile fatty acids (VFA)/
alkalinity ratio was found as 0.184. It confirms the stability of the reactor.
Keywords: alkalinity, anaerobic treatment, biogas, start-up, volatile fatty acids.
I. INTRODUCTION
Wastewater may be categorized as low-strength, medium strength, high strength and very high strength
based upon the concentration of chemical oxygen demand (COD), bio-chemical oxygen demand (BOD),
suspended solids (SS) and nutrients such as nitrogen (N) and phosphorus (P). Generally the stronger the
effluent, the greater the investment in infrastructure and energy inputs required to achieve environmentally
sound treatment objective [1]. Several techniques are already available to attain the goals of environmental
protection and resource conservation. In the case of wastewater treatment, combination of different methods can be
used, viz. physical, chemical and biological. The last method, which constitutes the main units of wastewater
treatment plants, can be divided into two classes such as aerobic and anaerobic [2].
Aerobic process, which are widely used for the treatment of wastewater have two distinct disadvantages
like high energy requirement and excess sludge production, which require handling, treatment and disposal [3]. In
contrast, anaerobic processes generate energy in the form of biogas, and produce sludge in significantly lower
amounts than those resulting from aerobic systems. In the absence of molecular oxygen anaerobic process
convert organic material into methane, a fuel that can yield a net energy gain process operation. Anaerobic
processes have gained popularity over the past 20 years. It have been already applied successfully for the
treatment of different type of wastewater and geared mainly towards high concentrated soluble wastewater [4].
The most common reactor type used for anaerobic digestion of wastewater is the continuously stirred tank
reactors (CSTR). The main problem of this reactor is the removal of active biomass from the system that leads
to long retention time. It has been overcome by immobilizing the active biomass, referred as high rate systems.
A typical such reactor is upflow anaerobic sludge blanket (UASB) reactor [5].
In the UASB process, wastewater flows through bed (granular or flocculent), where different physical
and biochemical mechanisms act in order to retain and biodegrade organic substances. Solubilisation and
hydrolysis of SS is a slow process accomplished by extracellular enzymes excreted by acidogenic bacteria.
Readily biodegradable substances are quickly acidified and then converted into methane and other biogas
components [6]. The great success of the UASB process lies in its capability to retain a high concentration of
active sludge due to granule formation. It allows the application of high loading rates and maintenance of high
solids retention time at a low hydraulic retention time (HRT) so that efficient treatment can be carried out in a
temperature climate as well [7].
Low-strength wastewater such as domestic sewage (COD concentration 500-1000 mg/L), are at
present treated mainly aerobically. However, due to considerable operation and maintenance cost, studies are
being conducted to develop economically more attractive high rate anaerobic systems [8]. UASB reactor system
is facing a challenge in the treatment of low-strength wastewater. Nevertheless, more efficient anaerobic

Start-up of an upflow anaerobic sludge blanket reactor treating low-strength wastewater inoculated
systems have been developed and they are being successfully applied for treatment of low-strength wastewater
such as domestic wastewater, particularly under tropical conditions where artificial heating can be avoided [9].
The production and quality of granular/flocculent sludge is influenced by the composition of wastewater, reactor
design and technological condition. Moreover, the formation of granular/flocculent sludge with good settling
characteristics and activity is also a critical factor in dealing with low-strength wastewater [10]. In general, the
operational conditions induce an ecological selection that leads to the formation of microbial aggregates, the so-
called ‘granules’ [11]. Granules are the best inoculum for the start-up of UASB reactors due to their high
specific activity and settleability. However, they are not always available. Consequently, very often the start-up
process and the tentative granule formation have to be performed with an inoculum of diffuse sludge collected
in conventional anaerobic digesters. There is lack of data on the operating conditions during the start-up of a
UASB reactor that uses diffuse sludge as the inoculum. On the contrary, when treating wastewaters with COD
values lower than 1500 mg/L, which are low- strength wastewater in the anaerobic treatment field, the start-up
of UASB reactors is difficult and thus much less full-scale reactors are in operation under these conditions than
in the case of medium and high strength effluents [9].
Keeping this in mind, the experimental work here described was focused on the anaerobic treatment of
low-strength wastewater under tropical conditions. The main objective of this work was to look into the start-up
process performance of a lab scale UASB reactor inoculated with digested slurry, which was collected from an
active biogas plant. The operational parameters such as COD, upflow velocity, total alkalinity, volatile fatty
acids (VFA) concentration, total suspended solids (TSS), volatile suspended solids (VSS) and pH were
investigated. In addition to that, biogas generation and COD removal efficiency were also calculated.
II. METHODS
2.1 Experimental set-up
UASB reactor employed in this study was fabricated using transparent plexiglass tube of 0.1 m
diameter vertical cylindrical shape with a total volume of 11.78 L. The reactor essentially had an internal
effective working volume of 9.97 L and the remaining volume of 1.81 L was kept for gas liquid solid (GLS)
separation arrangement. The overall height of the reactor was 1.5 m and effective height being kept as 1.27m.
The reactor had one influent port at the bottom, one effluent port and five sampling ports (Figure. 1). The GLS
separator attached at the top of the reactor was essentially a set of two inverted cones with intermediate overlap
to facilitate effective biogas collection. Gas was collected from the outer inverted cone as well as below the dead
corners of the inner cone. The treated effluent collected by a pipe attached above the GLS separator was
connected to a water seal to prevent the escape of gas. A peristaltic pump (Miclins PP 15) was used for feeding
wastewater as uniformly as possible over the reactor bottom, passes through the sludge bed in the reactor. The
reactor was operated at mesophilic temperature (24 - 35°C).
2.2 Synthetic wastewater preparation
In this study, synthetic wastewater was prepared, in which organic COD was provided by sucrose and
dried milk powder. About 1000 mg/L of COD contains 750 mg of sucrose and 250 mg of dried milk powder.
The ratio of soluble to particulate COD in the feed was around 80:20. The synthetic wastewater was then
neutralized to a value of pH ~ 7.2 by using sodium hydroxide (NaOH). Urea and KH2
PO4
were added as sources
of nitrogen and phosphorus respectively to maintain COD: N: P ratio. A ratio of 300:5:1 was maintained as
COD: N: P during start-up period [12]. The pH in the feed tank was adjusted to a value ~ 7 after every 10 -12
hours. The fresh feed was prepared daily. Trace elements like ferric chloride, copper sulfate, magnesium
chloride, calcium chloride were added appropriately.
2.3 Seed sludge
Digested slurry was collected from an active biogas plant located at Faculty of Agriculture, Annamalai
University, Annamalai Nagar. After analyzing the concentration of total suspended solids (TSS) and volatile
suspended solids (VSS), this sludge was used as seed. Initially about 50% reactor volume was filled up with this
active sludge.

1. Feed inlet, 2. Peristaltic pump, 3. Effluent outlet,
4. Biogas collection, 5. Gas measurement unit,
6. Sampling ports, 7. Effluent collecting tank,
8. 10 cm diameter and 150 cm tall plexiglass tube,
9. Gas liquid solid separation system
Figure1. Schematic diagram of UASB reactor showing experimental setup
2.4 Analysis
During the operation of the reactor, temperature, sludge bed height and biogas production were
monitored and recorded daily. Feed and treated effluent was taken for the analysis of COD, pH, total alkalinity
and VFA on daily basis. TSS and VSS were determined once in a week. Liquid samples were collected from the
influent, effluent and sampling ports along the reactor. Quality control was ensured using standards as well as
duplicates while performing analysis. All the parameters were determined according to standard method for the
examination of water and wastewater [13]. COD was determined by open reflux method. VFA was determined
by distillation method and measured as acetic acid per liter. Biogas evolved was collected by water displacement
method. The water in the gas collector was acidified with sulfuric acid to reduce the solubility of one of the
constituents of biogas carbon dioxide (CO2), for optimum measurement of biogas volume.
III. RESULTS AND DISCUSSION
3.1 Characteristic of seed sludge and inoculation
Initially the seed sludge (digested slurry) had a TSS content of about 70,100 mg/L and VSS content of
about 28,000 mg/L. Similar type of sludge was used as seed sludge for treating sago wastewater in a Hybrid
UASB reactor [14]. Lettinga and Hulshoff Pol [15] revealed that in order to avoid excessive sludge washout, the
amount of seed sludge must be small enough to maintain the sludge bed within the reactor upon increasing the
space-loading rate, and large enough to prevent a needless delay of the reactor start up. They also suggested,
when using thick digested slurry of relatively low specific methanogenic activity, 12 to 15 kg VSS/ m3
sludge
can be used because expansion washout will be limited in that case. While using s sludge with higher specific

activity, the amount of seed sludge should not exceed approximately 6 kg VSS/ m3
, because more seed sludge
will be washed out due to the occurrence of sludge bed expansion. Hence, initially about 50% reactor volume
was filled up with this active sludge having VSS concentration of about 28,000 mg/L. Accordingly the VSS
concentration in the entire reactor at the start-up was about 14,000 mg/L (14 kg VSS/m3
). Hickey et. al., [16]
recommended Sludge with 40% reactor volume might be enough to accelerate start-up.
3.2. Reactor Start-up
The start-up period is considered as the period taken for stable operation to be achieved. This is a
crucial step for the stable operation of the UASB and other anaerobic reactors, at a designed organic loading rate
(OLR). One of the main points stressed is the inoculation with high quality methanogenic sludge. Sludge of
poor quality can also be used [17]. If self-inoculation can be effected, the start-up periods can go up to or
beyond 6 months and other complications could arise [4]. In addition, operating temperature is prominent during
start-up. In this work, the UASB reactor after seeding was operated at a temperature between 240
C and 350
C.
This comes under the mesophilic range (200
C to 450
C). Fluctuations within this range were not enough to cause
any real impact on performance and in terms of reaction kinetics [18].
To begin with, the sludge has a VSS / TSS ratio of about 0.40. Poor generation of biogas at the initial
stage of start-up shows that sludge activity was initially low. At the same time, feed concentration of less than
1000 mg/L of COD helped in controlling the excessive volatile fatty acids (VFA) formation and guarded against
the excessive pH drop. An inhibition of methanogenesis generally results in an increase of VFA concentration
and a sudden drop in pH [19]. The upflow velocity is also an important operational parameter in anaerobic
digesters. It maintains the mixing and hydraulic retention time of the substrate and biomass [20]. The upflow
velocity maintained initially was low of about 0.106 m/h. No recirculation was employed to allow the lighter
fraction of biomass to wash out from the reactor. The upflow velocity maintained during start-up phase is
presented in Figure. 2. Accordingly, the OLR maintained during start-up phase is presented in Figure. 3.
COD removal rate during first four weeks were low in the range of 30-35% (based on total influent
COD and total effluent COD) (Figure. 3). The low removal efficiency during start-up phase can possibly be
attributed to the presence of unadapted seed sludge [10].Sufficient upflow velocities were maintained in the
reactor, in order to facilitate sludge blanket formation offering higher area for contact between sludge and
wastewaters. During period of 30 to 40 days higher upflow velocity of 0.128 m/h was tried (Figure. 2). This
upflow velocity was preferred to permit the wash out of voluminous (poorly settling) sludge. This amounted to
higher OLR of more than 2.1 kg COD/m
3
d (Figure. 3). However, these operating parameters made start-up
operation unstable as evidenced by increase in VFA concentration in the effluent. Hence, the upflow velocity
had to be controlled to a value of about 0.102 m/h after day 44 (Figure.2).
Many researchers suggest that the hydraulic retention times (HRTs) should not be allowed to less than
6 h and it should be less than 18 h during start-up period to treat any type of wastewater. At lower HRT, the
possibility of washout is more prominent. This makes it difficult to maintain the effective number of useful
microorganisms in the system [21]. For this reason, after day 3, the HRTs were varied between 10 h and 18 h
during start-up period.
Figure 2. Feed, effluent COD concentrations and upflow velocity maintained during start-up period
0
200
400
600
800
1000
1200
0 10 20 30 40 50 60 70 80 90
Time,days
COD,mg/L
0.000
0.050
0.100
0.150
0.200
0.250
Upflowvelocitym/h
COD mg/L Influent COD mg/L Effluent
Upflow velocity m/h

Figure 3. Organic loading rate maintained and % COD removal during start-up period
As discussed earlier during start-up period COD: N: P ratio was maintained as 300:5:1. N and P
supplementation was necessary characteristics. COD: N: P should be at least 300:5:1 for efficient rapid start-up
[12], [22]. Likewise, neutralization by alkali to a value of pH 7.0 - 7.2 was also necessary, particularly during
start-up. In addition, optimal methanogenesis is known to take place at a neutral pH. The well-being of a reactor
can often be judged by the examination of pH value of influent and the treated effluent. Variations in pH of the
influent and effluent are presented in Figure. 4. The value of pH slightly fell during the initial days of operation,
indicates the prevalence of acid fermentation over methanogens. However, after the initial drop, pH of the
treated effluent was in the range of 7.2 - 8.3. It indicates healthy anaerobic environment, which is favorable for
methanogenic organisms [23].
Figure 4. Feed and effluent pH during reactor start-up period
0
10
20
30
40
50
60
70
80
90
100
0 10 20 30 40 50 60 70 80 90
Time, days
%CODremoval
0.000
0.500
1.000
1.500
2.000
2.500
3.000
3.500
4.000
4.500
5.000
OLR,KgCOD/m3d
% COD Removal
OLR, kg COD/ m3 d

0
50
100
150
200
250
300
350
400
0 10 20 30 40 50 60 70 80 90
Time, days
VFA,Alkalinity,mg/L
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
VFA/alkalinityratio
Alkalinity mg/L Effluent
VFA mg/L Effluent
VFA/alkalinity ratio
Total alkalinity, VFA concentration and pH are effective indicators of the stability of the
reactor and of the correct balance between acidification and methanogenesis. The acidogenic element of the
overall digestion processes will generate acids, but under normal conditions this would be buffered by the
alkalinity in the reactor (expressed as calcium carbonate). Alkalinity may also be caused due to evolution of
CO2, during decomposition of organic matter [24]. In an anaerobic digester, a bicarbonate alkalinity (as CaCO3)
of about 1000 to 3000 mg/L was required for stable operation [25]. Disparity to this, Van Haandel and Lettinga
[26] shows an average alkalinity with a range of 220-390 mg/L for the successful operation of a UASB reactor
over 200 days. On this basis, sufficient alkalinity was available in the reactor. The alkalinity value of the treated
effluent considerably increases as the reactor progressing towards steady state conditions (Fig. 5). Methogenisis,
in particular is known to become unstable when the VFA / alkalinity ratio value is above 0.3 [27], [28]. From
the beginning of the experiment, the effluent VFA values were less than 225 mg/L. The bicarbonate ion
concentration or bicarbonate alkalinity is approximately equivalent to the total alkalinity for most wastes when
the volatile acid concentration is very low [29]. On this aspect, total alkalinity measured during the entire period
of operation represents bicarbonate alkalinity. In this work, VFA/alkalinity ratio after 40 days was 0.5. As the
reactor progresses towards stabilization, the ratio gradually decreases. At the end of start-up period, the VFA/
alkalinity ratio was found as 0.184 (Fig. 5). Most researchers revealed that a value of greater than 0.40 for this
ratio indicate reactor instability.
The VFA concentration (measured as acetic acid) in the effluent during first 30 days was in the range
of 225-170 mg/L from that it fell down to 108 mg/L on day 50. Higher levels of VFA in the wastewaters during
the initial phases of operation indicate the prevalence of acid fermentation [26]. Subsequently, the VFA in the
effluent decreased and was in the range of 80 to 59 mg/L towards the end of start-up period. It indicates healthy
anaerobic environment and satisfactory methanogenic activity. The overall performance of the reactor during
the start-up was more than satisfactory. The selection of seed material plays a crucial role in minimizing time
required for initial bioflim establishment [30], [31]. It was likely that the slurry collected from an active biogas
plant and used as a seed had sufficient numbers of physiologically active microorganisms.
The biogas production up to 20 days was low, less than 1.4 L/d. Beyond day 48, however as soon as
VFA concentration was controlled to less than 110 mg/L, the gas production as well as COD conversion
efficiencies started showing marked improvement. By day 84, the gas production rate reached a value of about
4.92 L/d (Figure. 6) and the VFA in the effluent had reduced to less than 60 mg/L (Figure. 5). This shows that
the process of hydrolysis and acidification of the organic matter took place in proper condition.
Figure 5. Effluent VFA, Alkalinity and VFA / Alkalinity ratio during start-up period.
It was assumed that the reactor had reached steady state condition merely 78 days after inoculation.
This is because the results of effluent COD presented the same pattern i.e. approximately the same average COD
concentrations and the same range of variations during the whole operation period after 78 days. There was no

recycling of sludge during entire operation. Effluent recycle was not necessary to fluidize the sludge bed as
sufficient contact between the wastewater and sludge is guaranteed even at low organic loading rate in UASB
reactor [32]. The start-up period for the reactor was completed in 84 days, as the VFA level in the effluent
stabilized gradually. The pH value of the treated effluent and increase in total COD removal efficiency (90.80%)
indicates good start-up.
Determination of VSS/TSS ratio gives correlation to the biomass growth and its quality. The VSS/TSS
ratios based on the average TSS and VSS concentrations in the reactor along with substrate utilization was
observed at different periods of operation. The biomass concentration profiles were obtained by the TSS and
VSS concentrations taken from sampling ports at various heights of the reactor (data not shown). At the time of
reactor startup, VSS/TSS was 0.40. There was a steady increase in VSS/TSS ratio throughout the operation.
The VSS/TSS ratio increased up to 0.64 by day 50 further increased to 0.80 on day 78. Initially the sludge bed
height inside the reactor was 0.70 cm. On day 50 and day 78, it was 0.40m and 0.45m respectively. However,
the experiment with the sludge morphological study was conducted at the end of the operational period.
Figure 6. Biogas production during start-up period
IV. CONCLUSIONS
The attributes gathered from this study points out the feasibility of UASB reactor for the treatment of
low-strength wastewater with COD values lower than 1000 mg/L, inoculated with non-granular sludge. The
development of sludge bed was observed during this start-up phase. Decrease in VFA values, increase in pH
values and alkalinity of the effluent, increase in biogas production rate and COD removal efficiency suggests
that phase separation, process of hydrolysis and acidification of the organic matter took place in proper
condition. It can be concluded that UASB reactor, inoculated by digested slurry requires 84 days for start-up and
may be a robust design for treating low-strength wastewater. Further investigation should be carried out for a
longer duration by varying OLR and HRTs to study the steady state and shock load performance. Effect of
variation in influent parameters on sludge activity and characteristics should be further studied.
V. Acknowledgement
One of the authors (K.R.Venkatesh) is thankful to the authorities of the Annamali University for
providing necessary facilities to carry out his research work.
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